CA1325770C - Adsorber module for whole blood treatment and an adsorber apparatus containing the adsorber module - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
A novel adsorber module for whole blood treatment is disclosed, which comprises a casing provided with a blood introduction means and a blood withdrawal means and a bun-dle of a plurality of porous hollow fibers disposed in the casing and disposed between and fluid-tightly connected at end portions thereof to the blood introduction means and the blood withdrawal means, wherein each porous hollow fiber comprises a membranous porous resin matrix having pores which open at least at the inner wall of the hollow fiber and a plurality of ligands linked to the overall surface, including the walls of open pores, of the porous resin matrix. The adsorber module can easily be con-structed into an adsorber apparatus which can be practical-ly employed for treatment of whole blood. With this appa-ratus, whole blood can be effectively, efficiently treated without the danger of blood coagulation and hollow clogg-ing, whereby the malignant components of the whole blood can be effectively removed by adsorption on the ligands.

Description

~ 132577~

Backqround of The Invention Field of The Invention The present invention relates to an adsorber module for whole blood treatment and an adsorber apparatus containing the adsorber module. More particularly, the present invention is concerned with an adsorber module for whole blood treatment comprising a casing provided with a blood introduc-tion means and a blood withdrawal means and a plurality of porous hollow fibers accommodated in the casing, wherein each porous hollow fiber comprlses a membranous porous resin matrix having pores which open at least at the inner wall of the hollow fiber and a plurality of ligands linked to the overall surface of the porous resin matrix.
Also, the present invention is concerned with an adsorber apparatus comprising the above mentioned adsorber module, a blood introducing passage means having one end fluid-tightly connected to the blood introduction of the module and a blood withdrawing passage means having one end fluid-tightly connected to the blood withdrawal means of the module.
By the use of the adsorber module and apparatus accordlng to the present invention, it is feasible to accomplish effective, efficlent treatment of ~.,~

" ~ -., ~ ~ . ;
.. . . , . , :
, ~ ' ~ . ,' ' .

~" 132577~

whole blood without the danger of blood coagulation and hollow clogging so that the malignant components of blood can be effectively removed by adsorption~
Discussion of Related Art In recent years, as a result of the remarkable progress of medicine (especially internal medicine), hematology, immunology, clinlcal inspection, etc., various malignant components have been isolated from patient blood and plasma and identified, which are believed to have close relation with the cause and advance of diseases. For example, such malignant components include autoantibodies and immune complexes against autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, hyperlipemea and the like. Also, such malignant components include low or very low density lipo-protein against familial hyperlipidemia and sub-stances of middle or low molecular weights which are - increased by liver diseases.
Accordingly~ efforts have been made in the art for developing an advantageous method by which the above-mentioned malignant component can be effec-tively removed from whole blood to thereby relieve or entirely cure the diseases caused by the compo-nent.

... - : ~ , , ., . ~ . .

~ 132~77~
.

The proposals which have hitherto been made ln line with the above-mentioned efforts are classiEied into (1) the one in which use is made of an active carbon or an active carbon having the surface there-of covered with a hydrophilic polymer material to remove the malignant component of plasma from whole blood, (2) the one in which whole blood is separated into a blood cell portion and a plasma portion, fol-lowed by removal of the malignant component from the -10 plasma portion using an adsorbent (see, for example, Japanese Patent Application Publicatlon Specifica-tion No. 55-22107/1980), and (3) the one in which the plasma separated from whole blood according to a method as mentioned in item (2) above is passed through a filter so as to remove the malignant com-ponent of high molecular weight from the plasma (see, for example, Japanese Patent Application Publication Specification No. 63-62/1988).
However, the proposals of items (2) and (3) above have disadvantages in that since separation of plasma from whole blood must be performed prior to adsorption or filtration, complicated apparatus is necessary and the operation is not easily accom-plishedO Moreover, in these proposals, since the priming volume (total of the inner volumes of ~ 132577~

apparatus components inclusive of a plasma sepa-rator, an adsorber or filter and conduits which are filled with blood) has inevitably been large, the volume of the body fluid taken from a patient has also been large to the detriment of the patient.
The proposals o item (1) above permit simpler apparatus and easier operatlon than those employed in the proposals in items (2) and (3) above, since whole blood treatment is performed without plasma separation. However, active carbon has poor selec-tivity for adsorption. Further, the pore si~e of active carbon is so small that the malignant compo-nent of the plasma cannot be adsorbed to a desirable extent.
In addition to the above-described proposals, more recently, it has been proposed to use a flat membrane in which the wall surface (disposed on the outer side relative to the blood path) has bonded thereto an antibody, an antigen t an enzyme or the like to remove the mallgnant component of plasma, such as an antigen, an antibody, an enzyme, a proteln, etc. In this connection, reference is made for example, to PCT Patent Application Publication No. WO 80/02805. However, this proposal has the disadvantage that the adsorbing capacity of the - .~.,.,, .. , :

.. ~ , ,~ : . ~, . .

bf' iL32!~i7PI~

hollow fiber is poor. Moreover, since an unstable biosubstance is used as a ligand in this proposal, sterilization is likely to cause the ligand to have a disadvantageously lowered activity or to cause the ligand to be detached from the flber, thereby bring-ing about serious problems with respect to safety.
Summary of The Invention With a view toward developing an effective device for removing the malignant component of whole blood which is free from the above-described draw-backs of the prior art, the present inventors have made extensive and intensive studies. As a result, it has unexpectedly been found that a desired device or apparatus for blood treatment can be obtained by the use of porous hollow fibers each comprising a membranous porous resin matrix having pores which open at least at the inner surface of each hollow fiber and a plurality of ligands linked or attached to the overall surface of the porous resin matrix, in which the porous hollow fiber is adapted to allow whole blood to flow within the hollow thereof along the length of the fiber while causing the plasma of the whole blood to contact at least the inner wall surface, including the open pore walls, of each hollow fibert so that the malignant component of the ,, , , . ,. ........ - .
, . . . . .

~ 13~5770 plasma is adsorbed on the hollow fiber by the inter-action between the ligand and the malignant compo-nent. Based on this novel finding, the present invention has been completed.
It is, therefore, an object of the present invention to provide an adsorber module by which it is easy to accomplish effect~ve, efficient removal of the malignant component of whole blood without the danger of blood coagulation and hollow clogging so that the malignant components of the blood can be effectively removed by adsorption.
It is another object of the present invention to provide an adsorber module which is not adversely affected by sterilization and can be used in whole blood treatment with safety.
It is a further ob~ect of the present invent~on to provide an adsorber apparatus containing an adsorber of the above character in which the priming volume is small.
~o It is still a further ob~ect oE the present invention to provide an adsorber apparatus which is simple is construction and easy in operation.
The foregoing and other objectsl features and advantages of the present invention will be apparent to those skllled in the art from the fo11owing :
., .

:
~?
.

~ 13~577~

detailed description and appended claims taken in connection with the accompanying drawings.
Brief Description of The Drawinqs In the dxawings:
Fig. 1 is a diagrammatic cross-sectional view of one form of an adsorber module according to the present invention;
Flg. 2 is an enlarged, diagrammatic cross~
sectional view of a portion of a preferred form of hollow fiber used in the present invention, illust-rating a plurality of ligands linked to the porous resin matrix of the hollow fiber~
Fig. 3 is a diagrammatic view of one form of an adsorber apparatus according to the present invention;
Fig. 4 is a diagrammatic view of another form of an adsorber apparatus according to the present invention; and Fig. 5 is a diagrammatic view of a comparative adsorber apparatus.
In Figs. 1 through 5, like parts or portions are designated by like numerals.
Detailed Description of The Invention In one aspect of the present invention, there !, 25 is provided an adsorber module for whole blood ~ - 8 -., .
:

1 3 2 ~ 7 7 0 treatment comprising:
a casing provided with a blood introduction means and a blood withdrawal means, and a plurality of porous hollow fibers substan-tially equal in length arranged subskantially ln parallel relationship and bonded together at both end portions thereof to fo~m a bundle, each porous hollow fiber of said bundle having openings at both terminal ends thereof, said bundle being dlsposed in said casing along the length of said casing, said both end portions of the hollow fibers of said bundle being fluid-tightly connected to said blood introduction means and said blood withdrawal means, respectively, thereby establishiny communica-tion between said blood introduction means and said blood withdrawal means through said bundle of hollow fibers, each porous hollow fiber comprising a membra-nous porous resin matrix having pores which open at least at the inner wall o~ the hollow fiber and a plurality of ligands attached to the overall surface of said porous resin matrix t said ligands being capable of interacting with a substance to be adsorbed.

_ g ;

- ` ' ' ~ 132~770 Referring now to Fig. 1, there is shown a schematic cross-sectional view of one form of adsorber module of the present invention which has opening 4 in the wall of the casing. The adsorber module comprises a casing provided with blood intro-duction means 2 and blood withdrawal means 3; and a plurality of porous hollow fibers 1 substantially equal in length which are arranged substantially in parallel relationship and bonded together at both end portions thereof to form a bundle. Each porous hollow fiber 1 of the bundle has openings at both terminal ends thereof~ The bundle is accommodated in the casing along the length of the casing. The both end portions of hollow fibers 1 of the bundle are fluid-tightly connected to blood introduction means 2 and blood withdrawal means 3, respectively, thereby establishing communication between blood introduction means 2 and blood withdrawal means 3 through the bundle of hollow fibers 1.
The porous hollow fiber employed in the adsorber module of the present invention comprises a membranous porous resin matrix having pores which open at least at the inner wall of the hollow fiber and a plurality of ligands linked or attached to the overall surface of the porous resin matrix. The - .- `,~' ' ' ', P ~32~77~
terminology "overall surface" as used herein means all o the inner and outer surfaces of the hollow fiber membrane and the surfaces defined by the walls of open pores present within the membranous porous resin matrix.
In Fig. 2~ there is shown a schematic cross-sectional view of a preferred form of porous hollow fiber used in the present invention. Porous hollow fiber 1 shown in Fig. 1 comprises membranous porous resin matrix lalso designated numeral I in Fig. 2) and a plurality of ligands 8 linked to the overall surface of the porous resin matrix. The resin matrix 1 has through-paths running between both surfaces 5 and 6 of the resin matrix. As mentioned above, ligands 8 are linked or attached to the overall surface of matrix 1, that is, lnner wall surface 5, outer wall surface 6 and pore wall surface 7~
In the present invention, the pores in the resin matrix do not necessarily form through-paths running through the thickness of the resin matrix as long as the pores open at least at the inner wall of the hollow fiber, however, from the viewpoint of attaining efficient, efective removal of the malig-nant component of whole blood, it is preferred -that :. ...

~ 132~77~

the membranous porous resin matrix of each hollow fiber have pores therewithin and openings on both surfaces thereof~ which pores cooperate with the openings to form through-paths running between both the surfaces of the resin matrix.
The average pore diameter of the porous resin matrix is not particularly limited in the present invention. The pore diameter is generally selected taking into account the size and shape of the malig-nant component to be removed by the porous hollow fiber membrane. It is preferred that the hollow fiber have a pore diameter at which the plasma con-taining the malignant component is freely passed through the membrane so that the inner wall surfaces o~ the pores are so large as to permit unlimited contact of the plasma containing a malignant compo-nent with the ligands linked to the porous resin matrix. The average pore diameter is determined as followsO The pore diameter and pore volume of the membrane are measured by means of a mercury porosi-meter. The logarithm of the diameter is plotted as '!
the abscissa and the pore volume is plotted as the ordinate to give a pore diameter distribution curve.
Thus, the total pore volume is defined by the abscissa and the pore diameter distribution curve.

.

~ ~ ~2~;P~7~

A vertical line can be drawn in parallel to the ordinate so that the total pore volume ~s halved.
The value of the pore diameter on the abscissa at its po1nt crossed by the above-mentioned vertical line is referred to as the "average pore diameter"O
The average pore diameter is generally in the range of from 0.005 to 3 ~m, preferably from 0.01 to 2 ~mr more preferably from 0.02 to 1 ~m.
The area of the overall surface of the porous resin matrlx is not critical in the present inven-tion. However, the area of the overall surface is generally at least 5 m2/g, preferably at least 10 m2/g, more preferably at least 15 m2/g. The area of the overall surface is determined as follows~
The nitrogen gas adsorption amount of the matrix is measured using a BET ~ Sr Brunauer - PH~ Emmett - E.
Taylor) type surface area measuring apparatus, and the overall surface is determined from the nitrogen gas adsorption amount according to the customary one-point method.
In the present invention, it is preferred that the porous hollow fibers have an average effective length (L mm~ and an average inner diameter (D mm) which satisfy the inequality:
L/D~ ~mm~1) ~' 2000.

- 13 _ : . - , - .
: . ,~ :
~, .... , , . -,. . . , ,. :- ~. .

~ ~32~77~

The average effective length ls defined as an average of the lengths of the porous hollow fibers minus the lengths of both end portions of the porous hollow fibers which bo~h end portions are bonded together and fluid-tightly connected to blood intro-duckion means 2 and blood withdrawal means 3, respectively.
In the adsorber module of the present inven-tion, each porous hollow fiber is adapted to allow whole blood to rlow within the hollow thereof along the length of the fiber while causing the blood to be contacted with the wall surface of each hollow fiber, so that the malignant components of the blood is adsorbed on the hollow fiber by the interaction between the ligands and the malignant components.
Since a large surface is effectively utilized for whole blood to be contacted with the ligands linked or attached to surface of the porous resin matrix, effective, efficient removal of the malignant compo-nents of whole blood can be attainedO
When the membranous porous resin matrix of each hollow fiber have pores therewithin and openings on both surfaces thereof, which pores cooperate with the openings to form through-paths running between both the surfaces of the resin matrix/ the volume .: . , ~ ... . ~

~ ~L32~77~

ratio of erythrocyte to whole blood (hematocrit) is generally higher at portions of the hollow of the porous hollow fiber which are on the side of the blood withdrawal means than at portions of the hollow of the porous hollow fiber which are on the side of the blood introduction means. This is attributed to permeation of the plasma through the wall of the hollow fiber from the inside of the fiber to the outside of the fiber. The above-mentioned increase in the volume ratio of erythrocyte brings about an increase in the viscos-ity of the blood flowing through the hollow of the porous hollow fiber. The increase in blood viscos-ity produces an advantageous pressure by which the above-mentioned permeation of blood plasma is promoted. On the other hand~ in the adsorber module on the side of the blood withdrawal means, plasma which has been passed through the hollow fiber membrane and has entered the interstices extending between the outer wall of the porous hollow fibers and the inner wall of the casing and between the outer walls of the individuall hollow fibers is returned to the hollow of the porous hollow fiber by the action of the increased pressure of the plasma in the outside of the hollow fiber. The plasma ... ..
: ~ . , -,:: ~ .. .
:; : - - :

~ .

~ ~32~77~

joins the blood havlng an increased content of erythrocyte in the hollow of the porous hollow fiber, and then the resultant blood is discharged from the adsorber module through the blood with-drawal means. When the plasma first passes from the inside to the outside of the hollow fiber through the hollow fiber wall membrane, a great part of the malignant component is adsorbed. And, when the plasma then passes from the outside to the inside of the hollow fiber through the hollow fiber wall membrane, the malignant component is almost completely removed.
The plasma having most the malignant component removed by first passing it from the inside to the outside of the hollow fiber through the wall membrane, may be discharged from the casing through at least one opening provided for withdrawing the plasma. The plasma discharged from the casing flows through a conduit connected to the opening, and then joins the blood flowing through a conduit which is connected to the blood withdrawal means of the casing~ as described later.
From the viewpoint of minimi~lng the priming volume as well as increasing the malignant compo-nent-separating efficiency by the passage of plasma :' ~ 16 -..

~ 132~770 through the hollow fiber membrane, it is preferred that the lnner diameter of the porous hollow fiber be as small as possible on the condition that blood can Elow through the hollow of the porous hollow fiber. Specifically, the inner diameter of the porous hollow fiber is preferably 100 ~m to 1,000 ~m, more preferably 150 ~m to 400 ~m.
In the present invention, it is preferred that the wall thickness of the porous hollow fiber be as large as possible as long as the efficiency for passing plasma through the hollow fiber membrane is not practically impaired. By increasing the wall thickness, the total surface area of the pore wall of the porous hollow fiber is increased. Specifi-cally, the wall thickness of the porous hollow fiber is preferably 10 ~m or more, more preferably 50 ~m to 500 ~m, and most preferably 50 ~m to 200 ~m.
The membranous porous resin matrix to be used for preparing the present porous hollow fiber may be prepared from a hydrophilic material, such as cellu-lose, a cellulose derivative, a water-insoluble polyvinyl alcohol and a copolymer o~ ethylene and vinyl alcohol or a hydrophobic material, such as a polyolefin (e.g., polyethylene or polypropylene), a polysulfone and a polytetrafluoroethylene. It ~ - 17 -..

'' , ~ ' :

~: -: ~: , .. :
:. .: .

~` ~32~70 should be noted that when a hydrophobic material is used, since an aqueous solution is not readily passed through a porous hollow fiber made of the hydrophobic material, it is generally necessary to render the surface of the porous hollow fiber hydro-philic, e~g., by coating the surface of the porous hollow fiber with a hydrophilic material or by subjecting the surface of the porous hollow fiber membrane to chemical treatment or plasma treatment.
From the viewpoint of facilitating the linking of the ligands which interact with the malignant component at the surface of the membranous porous resin matrix~ it is preferred that the matrix have functional groups to which the ligands are readily bonded, such as a hydroxyl group, an amino group, a carboxyl group and a thiol group~ However, since ligands can be linked to the surface of the porous hollow fiber by physical adsorption, embedding, insolubilizing precipitation onto the matrix surface and the like, the porous resin matrix does not necessarily have to possess may such functional groups.
There is no particular limitation with respect to the method for forming a porous hollow fiber.
That is, a porous hollow fiber can be formed by a ,, .

~' ., :,. :
.: : , . . ~ - . :,.
~ . r~ :
' " , ' ' ` :;' . ~
',',', : .'' , ' , ' ~'~. :
:,; ' ' ' : :, ' : . :.

~ ~32~7~

customary method, such as wet spinning, dry spin-ning, melt spinning or the like. It i5 preferred to prepare a porous hollow fiber by a stretching per-foration method in which a crystalline polymer is spun, e.g., by melt spinning, and subjected to cold stretching to cause cleavage among crystalline lamellae of the polymer and then subjected to hot stretching to attain an expansion of the cleavage.
This is because controlling of pore diameters of the porous resin matrix of hollow fiber is easy and because the mechanical strengths of the obtalned hollow fiber are high irrespective of the presence of a large number of pores. It is also preferred to prepare a porous hollow fiber from a polyolefin by the stretching perforation method from the viewpoint of the chemical resistance of the matrix of the hollow fiber during the reaction for linking ligands to the surface of the membranous porous resin matrix of hollow fiber.
The ligands may be linked to the surface of the membranous porous resin matrix by any of the custom-ary methods. For example, the ligands are bonded to the surface of the membranous porous resin matrix by, e.g., covalent bonding or ionic bonding. ~lso, the ligands may be bonded to the surface of the :

:, .. . .

::;; . .. . . : . , .
: . : ~ . :

: :,; - . . ., . - ~

~ ~3257~

porous matrix by physical adsorption, embedding, insolubilizing precipitation onto the matrix surface or the like. Of these methods, from the viewpoint of avoiding the elution of ligands, it is preferred to use a method in which the ligands are bonded to the surface of the matrix by covalent bonding.
Specifically, the conventional carrier activating method and the conventional ligand linking method, which are utilized in the preparation of an immobi-lized enzyme or in an affinity chromatography, may be employed. Representative examples o-f carrier activating methods include methods using a cyanogen halogenide, periodic acid, a crosslinlcing agent or an epoxide. In the carrier activating method, substitution and/or addition of active hydrogen-containing nucleophilic groups, such as an amino group, a hydroxyl group, a carboxyl group and a thiol group, to the reactive groups on the surface of the porous resin matrix is performed by chemical modification with, e.g., a cyanogen halogenide, such as CNBr. However, the carrier activating methods are not restricted to the above-mentioned methods.
"Ligand" used herein means a substance which selectively interacts with a malignant component to be adsorbed on the hollow fiber, which is contained : . . - :

: - . ~ . .-. : , : .: :
.

~32577~

in blood plasma.
Representative examples of ligands include an amino acid, a peptide, a protein, a glycoprotein, a monosaccharide, an oligosaccharide, a polysaccha-ride, a lipid, an antigenic substance, a nucleic acld, a nonprotein organic compound and an inorganic compound. Representative examples of pxoteins and glycoproteins include an antibody, a complement, a protein relating to blood coagulation and an enzy~e.
However, it is pre~erred that the ligands exhibit no or only weak antigenicity and toxicity even if it is eluted into blood, and that when the ligands are in contact with ~lood corpuscles, the ligands do not cause hemolysis of erythrocyte t sensitiæation of leukocyte or adherence and/or aggregation of plateletc In this connection, when a peptide, a protein and a glycoprotein have a low weight average molecular weight, they ha~e a low antigenicity. Therefore, with respect to a peptide, a protein and a glycoprotein, it is preferred that they have a low weight average molecular weight, although there is generally no particular restric-tion with respect to the molecular weight of the ligands. From these viewpoints, preferred are a peptide, a protein and a glycoprotein, each of which .

'"

~ :. : . :

~ 13~577~

has a preferred weight average molecular weight of not greater than 104; and a monosaccharide, an oligosaccharide, a polysaccharide, a lipid, a nucleic acid, a nonprotein organic compound (e.g., a polyanion), an inorganic compound, an amino acid and a peptide each having a weight average molecular weight of not greater than 103. The weight average molecular weight of the peptide, the protein and the glycoprotein are determined by measuring a sedimen-tation coefficient of the solute. That is, the sedimentation coefficient of the peptide, the protein or the glycoprotein is measured according to the sedimentation velocity method using an ultra-centrifuge, and the molecular weight is calculated by substituting the sedimentation coefficient measured for the s in the following Svedberg formula:
R.T.s Welght average molecular weight(Mw~ =
D(1-vP) ~o wherein ~ represents the gas constant, T represents the absolute temperature, s represents the sedimen-tation coefficient of a solute, D represents the diffusion coefficient of a solute, v represents the partial specific volume of a solute and p represents the density of a solvent.
.

.~ .
i ., : , . .:
~`' :' ~: . .
- - - ~

:, :

~ ~32~7~
Representative examples of ligands are as follows.
For the treatment of systemic lupus erythemato-sus, preferable use is made of a mononucleoside containing adenine, guanine, cytosine, uracil or thymine, an oligonucleoside or polynucleoside comprising such a monomucleoside; and a natural nucleic acid, such as DNA and RNA. These are used to remove an antl-nuclear antibody and an anti-DNA
antibody which are present in blood plasma.
Further, a basic compound, such as actinomycin D, may be used to remove DNA, R~A and ~NA which are present in blood plasma.
For the treatment of the so-called immune complex diseases~ such as chronic articular rheuma-tism, malignant tumor and systemic lupus erythema-tosus, a hydrophobic compound may be used to remove an immune complex.
For the treatment of diseases so-called auto-immune diseases, such as myasthenia, multiple sclerosis and chronic articular xheumatism, a hydro-phobic compound may be used to remove an autoanti-body.
For the treatment of hyperlipemia, use is preferably made o~ a polyanion, e.g., an acid poly-,. : ~, --..,...., . ,-:., : .. :
- . . . : .

'' '.:"': ~ ' ~ 1~2~77~

saccharide, such as heparin and dextran sulfate; a synthetic polyanionl such as polyvinyl sulfate and polyacrylic acid. These are used to remove a low density lipoprotein and a very low density lipo-protein.
The ligands to be used in the present invention are not restricted to the above-mentioned ones. The ligands may be used individually or in combination.
When the ligands are used in combination, the bundle to be incorporated in the present adsorber module may be comprised of hollow fibers of the same kind, each having different types of ligands or of di~ferent hollow fibers which are different in the type of ligand linked thereto.
With respect to a hydrophobic compound and a polyanion used as ligands, a more detailed explana-tion will be made as follows.
The hydrophobic compound to be used in the present invention is a compound having a solubility in physiological saline of not greater than 100 mmole/dl (at 25 C), prefexably not greater than 30 mmole/dl (at 25 C). When a compound having a solubility exceeding 100 mmole/dl in physiological saline is used, the compound is so hydrophilic that the compound has poor affinity for an autoantibody i.

- 24 _ ,R

`' . ~ ' ' ~ ' '' ' ' ' ' `~ ~'' . . . . . .
. . ., , ~

''' . ' " ' ' ,' ' .

~ ~32~770 and an immune complex, thereby leading to an extreme lowering in the adsorbing capacity of the adsorber module. Further, since such a compound has an aEfinity for an albumin which is hydrophilic, the albumin is likely to be non-selectively adsorbed on the porous hollow fiber. This is disadvantageous.
When a hydrophobic compound having at least one aromatic ring is used as the ligand, particularly preferable results can be obtainedO A hydrophobic compound having any type of aromatic ring may be advantageously used. However, it is preferred to use a hydrophobic compound having at least one aromatic ring selected from a benzene type aromatic ring, such as a benzene ring, a naphthalene ring and a phenanthrene ring; a nitrogen-containing six-membered ring, such as a pyridine ring, a quinoline ring, an acridine ring, an isoqulnoline ring and a phenanthridine ring; a nitrogen-containing Eive-membered ring, such as an indole ring~ a carbazole ring, an isoindole ring, an indolizene ring, a porphyrin ring and 2,3,2',3'-pyrrolopyrrole ring; a six-membered ring having 2 or more nitrogen atoms, such as a pyridazine ring, a pyrimidine ring, a sym-triazine ring, a sym-tetrazine ring, a quinazolin ring, a 1,5-naphthyridine ring, a pteridine ring and .~

, .

i, 132~7~70 a phenazine ring; a five-membered ring having 2 or more nitrogen atoms, such as a pyrazole ring, a 1,2,4-triazole ring, a 1,2,3-triazole ring, a tetrazole ring, a benzimidazole ring, an imidazole ring and a purine ring; an oxygen-contalning aromatic ring, such as a norharman ring, a per-imldine ring, a benzofuran ring, an isobenzofuran ring, and dibenzofuran ring; a sulfur-containing aromatic ring, such as a benzothiophene ring, a thienothiophene ring and a thiepine ring; an oxygen-containing heterocyclic aromatic ring, such as, an oxazole ring, an isooxyzole ringr a 1,2,5-oxadiazole ring and a benzoxazole ring; a sulfur-containing heterocyclic aromatic ring7 such as a thiazole ring, an isothiazole ring, a 1,3,4-thiadizaole ring and a benzothiazole ring; and derivatives thereof. Of these compounds, a compound having an indole ring, such as tryptamine, provides particularly preferred results. This is probably because the hydrophobic nature and rigidity of the molecule of such com-pounds promote the bonding of the compounds with an autoantibody and an immune complex.
It has been found that a hydrophobic amino acid and a derivative thereof which are practically harm-less and inexpensive are extremely advantageous to _ 26 -.

,:- ' , ' ., ~ ' ,'.. '' .. . . .
: - . . . .. :
: ,: . ' ~ ~3257~

remove an autoantibody and an immune complex. The hydrophobic amino acid and a derivative thereof preferably have a hydrophobic parameter of 1500 cal/mol or more, which hydrophobic parameter is defined by Tanford and Nozaki in Journal Of American Chemical Society 184, page 4240 (1962) and Journal Of Biological Chemistry 246, page 2211 (1971).
Also, the hydrophobic amino acid and a derivative thereof preferably have a solubility in physiolog-ical saline of 100 mmole/dl or less (at 25 C).
When the solubility exceed 100 mmole/dl, the hydro-phobic interaction between the hydrophobic amino acid and a derivative thereof and a malignant compo-nent becomes weak, thereby causing the adsorber module to have poor adsorbing ability.
Representative examples of hydrophobic amino acids and derivatives thereof include lysine, valine, leucine, tyrosine, phenylalanine, isoleucine, tryptophan and derivatives thereof.
Of these hydrophobic amino acids and deriva-tives thereof, phenylalanine, kryptophan and deriva-tives thereof are particularly preferred. There is no particular limitation with respect to the optical ~ isomerism of the hydrophiblic amino acid. That is, `~ 25 the hydrophobic amino acid may be dextrorotatory or _ 27 -... . ~......... .
.. , . . . ~ . .

P ~32577~

levorotatory.
The weight average molecular weight of the hydrophobic compound to be used in the present invention is preferably 10,000 or less, more prefer-ably 1,000 or less. The weight average molecular weight of the hydrophobic compound is determined in substantially the same manner as in the measurement of the weight avexage molecular weights of the peptide, the protein and the glycoprotein. When the hydrophobic compound has the above-mentioned prefe~-able weight average molecular weight, the hydro-phobic compound is readily linked to the membranous porous resin matrix of hollow fiber, and the hydro-phobic compound-linked porous hollow fiber has good storage stability as compared to that of a natural polymer, such as protein S ~weight average moleculax weight: 42,000). Even if a hydrophobic compound having a weight average molecular weight of 10,000 or less is eluted from a porous hollow fiber into blood, since the antigenicity of the hydxophobic compound is negligibly low, there arises no safety problem for a living body. The hydrophobic compound having a weight average molecular weight of 10,000 or less is not deactivated by sterilization.
The term "polyanion" used herein means a .. ., ,,, . ~, : . .

~..

1325~ ~

polymer which has a weight average molecular weight of 600 or more and which has functional groups capable of produciny a negative charge ln a body fluid, such as a sulfonic group, a carboxyl group and a phosphoric group. The weight average molec-ular weight of the polyanion is determined in substantially the same manner as in the measurement of the weight average molecular weights o~ the peptide, the protein and the glycoprotein. The polyanion is preferably in the form of a straight chain polymer so as to capture a malignant component effectively. The weight average molecular weight of the polyanion is preferably in the range of 600 to 107, more preferably 1,000 to 5 x 106, most prefer-ably 2,000 x 106. Since a low density lipoprotein and a very low density lipoprotein, which are malig-nant components, have a diameter as large as 2 x 10-4 to 8 x 10-4 ~m, it is desired to use a polyanion having the above-mentioned weight average molecular weight. When the polyanion has a weight average rnolecular weight of less than 600, the adsorbing capacity for a low density lipoprotein and a very low density lipoprotein is likely to be insuf-ficient. It is preferred that the polyanion have at least one of the above-mentioned functional groups '. ' ' ~ ~ :, : `

~`~; 132~70 capable of producing a negative charge in a body fluid, such as a sulfonic group, a carboxylic group and a phosphoric group per 300, more preferably per 200, most preferably per 50 to 150 in weight average molecular weight of the polyanionO This is because strong electrostatic interaction can be attained between the polyanion and a malignant component.
This weight average molecular weight of the poly-anion means a weight average molecular weight which includes the molecular weight of the functional groups.
Representative examples of polyanions include vinyl type synthetic polyanions, such as a poly-acrylic acid, a polymethacrylic acid, a polyvinyl sulfonic acid, a polyvinyl sulfuric acid, a poly-maleic acid, polyfumaric acid and derivatives thereof: styrene type synthetic polyanions, such as a poly(styrenesulfonic acid) and a poly(styrenephos-phoric acid); peptide type polyanions, such as a polyglutamic acid and a polyaspartic acid; nucleic acid type polyanions, such as a poly U and a poly A;
synthatic polyanions, such as a polyphosphoric acid ester, a poly-a-methylstyrenesulfonic acid and copolymers o~ styrene and methacrylic acid; and polysaccharide type polyanions; such as heparin, ;

. - 30 -. .
`~. ' . , :. ~

~ ~32~77~

dextran sulfate, chondroitin sulfate, alginic acid, pectin, hyaluronic acid and derivatlves thereof.
However, the polyanoin to be used in the present invention is by no means restricted to the above-mentioned specific examples.
The ligands capable of interacting with a malignant component are linked to the overall surface of a membranous porous resin matrix, prefer-ably with substantially uniform distribution.
However, in some portions, the ligands may be non-uniformly linked to the surface of the membranous porous resin matrix. Further, in the surface of a membranous porous resin matrix, there may be present some portions to which the ligands are not linked, as long as the presence of such portions cause little adverse effect on the adsorbing capacity for a malignant component.
The distribution of the ligands linked to the inner wall surface and the outer wall sur~ace of the membranous porous resin matrix can be observed by coloring the ligands according to the customary staining methodO The distribution of the ligands linked to the wall surface of the pores of the porous resin ma~rix can be observed by immersing the porous ~iber in an aqueous solution of a metal salt - , ~ ~2~7~

to form a salt of the ligand with the metal ut.iliz-ing the interackion between the ligand and the metal, cutting off a thin section from the resultant porous fiber and subjecting the thin section to observation using an X-ray micro analyzer to detect the metal which is present in the form of the salt of the ligand with the metal linked to the wall of the pores of the porous fibex.
When the inner surface of the porous hollow fiber is subjected to treatment for prevention of hollow clogging by platelets and/or treatment for preventlon of blood coagulation, more preferable results are obtained.
The porous hollow fibers are packed in a casing to prepare the adsober module for blood treatment according to the present invention by the use of a customary method, e~g., a method for preparing an artificial kidney.
The adsober module for whole blood treatment of the present invention can advantageously be constructed into an adsorber apparatus.
Accordingly, in another aspect of the present . invention, there is provided an adsorber apparatus for whole blood treatment comprisiny:
(a) an adsorber module comprising:
. ~

: . . .
/

:,,~ ~ . . ~ :., ~ ~ ~2~77~

a casing provided with a blood introduction means and a blood withdrawal means, and a plurality o~ porous hollow fibers substan-tially equal in length arranged substantially in parallel relationship and bonded together at both end portions thereof to form a bundle, each porous hollow fiber of said bundle having openings at both terminal ends thereof, the bundle being disposed in the casing along the length of the casing, the both end portions of the hollow fibers of the bundle being fluid-tightly connected to the blood introduction means and the blood withdrawal means, respectively, thereby establishing communica-tion between the blood introduction means and the blood withdrawal means through the bundle of hollow fibers, each porous hollow fiber comprising a mem-branous porous resin matrix having pores which open at least at the inner wall of the hollow fiber and a plurality of ligands attached to the overall surface of said poxous resin matrix, the ligands being capable of interacting with a substance to be adsorbed, ~b) a blood introducing passage means compris-. - 33 -.'`~ ' ., ,' , ~' ,' . . ' , ' ... ' , . ' ~ ~ 32577~

ing a first conduit having one end fluid-tightly connected to the blood introduction means of module (a), a second conduit having a blood inlet at one end thereof, and a blood transport means disposed between and fluid-tightly connected to the other ends of the first and second conduits; and ~c) a blood withdrawing passage means compris-ing a third conduit having one end fluid-tightly connected to the blood withdrawal means of module ~a) and having a blood outlet at the other end thereof.
In the adsorber apparatus~ preferably, the membranous porous resln matrix of each hollow fiber of the adsorber module may have pores therewithin and openings on both surface thereof, which pores cooperate with the openings to form through-paths running between both the surfaces of the xesin matrix. In this case, the casing of module (a) may further be provided with at least one opening for withdrawing the plasma which opening ls fluid-tightly connected to the blood withdrawing passage means through a bypass comprising a fourth conduit having one end fluid-tightly connected to the opening, a fifth conduit having one end fluid-tightly connected to the blood withdrawing passage at its intermediate ; - 34 _ .

: ~ . . .
.;, ~ , .~ . . .
..

.

~ i32~7~

position, and a plasma transport means disposed between and fluid-tightly connected to the other ends of the fourth and fifth conduits.
Referring to Fig. 3, there is shown one form of adsorber apparatus of the present invention. The apparatus comprises (a) adsorber module 13 having no opening (as indicated in Fig~ 1) in the casing for withdrawing the plasma; ~b) a blood introducing passage means comprising first conduit 12 having one end fluid-tightly connected to module 13 at its blood introduction means 2 (not shown), second conduit 10 having blood inlet 9 at one end thereof, and blood transport means 11 (e.g., pump) disposed between and fluid-tightly connected to the other ends of first and second conduits 12 and 10; and (c) a blood withdrawing passage means comprising third conduit 14 having one end fluid-tightly connected to module 13 at its blood withdrawal means 3 (not shown) and having blood outlet 15 at the other end thereof. Each porous hollow ~iber 1 of module 13 is adapted to allow whole blood, which is introduced from blood inlet 9 through the blood introducing passage, to ~low within the hollow thereof along the ` length of fiber 1 while causing the plasma of the ; 25 blood to be contacted at least inner wall sur~ace 5, ' ' `' ~ , : ' . : .

:: , . .
.

~ ~32~77~

including open pore walls, of each hollow fiber 1, so that the malignant component of the plasma is adsorbed on hollow fiber 1 by the interaction be-tween ligand 8 and the mallgnant component.
In operation, whole blood is introduced to the adsorber apparatus from the patient (donor) through blood inlet 9 and flowed to adsorber module 13 at a predetermined flow rate through conduits 10 and 12 by means of blood transport means 11 ~e.g., pump).
The whole blood is treated during the flowing through the hollow fibers so that the malignant components of the blood are adsorbed on the ligands attached to the hollow fiber membranes. The blood treated with the adsorber module 13 is flowed through conduit 14 and returnad to the donor through blood outlet 15. The whole blood from the donor is recycled through the same absorbex apparatus. The recycling is continued for a predetermined period of time. After the recycling purified~ the whole blood of the patient becomes pure~
It is preferred that the membranous porous resin matrix of each hollow fiber 1 of adsorber module 13 have pores therewithin and openings on both surfaces and thereof, which pores cooperate with the openings to form through-paths running be-.~ .

:: . :. , . :

::
:.: : ::
~ . : :
: . . .

S~ ~32~77a tween both surfaces and of the resin matrix. When such hollow fibers are used in adsorber module 13, during the flowing of the whole blood through the hollows of the hollow fibers, the plasma permeates through the wall of each hollow fiber from the inside of the fiber to the outside of the fiber.
The plasma which has been passed through the hollow fiber membrane and has entered the interstic~s extending between the outer wall of the porous hollow fibers and the inner wall of the casing and between the outer walls of the individual hollow fibers is returned to the hollow of the porous hollow fiber by the action of the increased pressure of the plasma in the outside of the hollow fiber.
This is advantageous from the viewpoint of attaining a greater contact area between the whole blood and the ligand.
Refexring to Fig~ 4, there is shown another form of the adsorber apparatus of the present inven-tion. In adsorber module 13, as described with reference to Fig~ 2, the membranous porous resin matrix of each hollow fiber 1 of adsorber module 13 has pores 7 therewithin and openings on both surfaces 5 and 6 thereof, which pores 7 cooperate with the openings to form through-paths running be-, . : :.... :- - ~, :: ~, ~ 2 ~ 7 7 0 tween both surfaces 5 and 6 of the resin matrix.
Therefore, as described above, when the whole blood is allowed to flow within the hollows along fibers 1, the plasma of the blood is caused to be selec-tively passed from the inside to the outside and, in turn, from the outside to the inside of each hollow fiber 1 reciprocally through the walls of hollow fibers 1. In the apparatus of Fig. 4, the casing of adsorber module 13 is further provided with at least one opening for withdrawing the plasma. The opening is fluid-tightly connected to blood withdrawing passage means (19 and 20) through a bypass compris-ing fourth conduit 16 having one end fluid-tightly connected to the opening, fifth conduit 18 having one end fluid-tightly connected to blood withdrawing passage (19 and 20) at its intermediate position, and plasma transport means 17 (e.g., pump) disposed between and fluid-tightly connected to the other ends of fourth and fifth conduits 16 and 18.
In operation, whole blood is introduced in the adsorber apparatus from blood inlet 9 and flowed to adsorber module 13 at a predetermined rate through conduits 10 and 12 by means of blood transport means 11 (e.g., pump). In adsorber module 13, the blood i5 allowed to flow through the hollows of hollow i .

, . . ..
. . : `
~: .

~ ~32577~

fibers 1~ and the plasma of the blood is allowed to pass through the membrane of each hollow fiber. The thus separated, purified plasma i5 flowed to conduit 20 through a bypass, i.e., through conduits 16 and 18 by means of blood transport means 17 (e g., pump~
at a ~low rate which is lower than the flow rate of the blood flowing from conduits 10 and 12 into adsorber module 13 by means of pump 11. On the other hand, the cell-enriched blood which has not been passed through the bypass is flowed to conduit 20 through conduit 19. In conduit 20, the purified plasma is combined with the cell-enriched blood and the resultant mixture is returned to the donor through blood outlet 15. The blood from the patient lS is recycled through the same adsorber apparatus.
The recycling is continued for a predetermined period of time. After the recycling, the whole blood of the patient becomes pure.
As described, the adsorber module of the present invention can easily be constructed into an adsorber apparatus which can be practically employed for treatment of whole blood. With this apparatus, whole blood can be effectively, efficiently treated without the danger of blood coagulation and hollow clogging, whereby the malignant components of the .

! ' .
': ' ~ ' , , ': " '" ' ' '', " . ~ '"' ' ' " ' """ ''. ` . .

~ ~32~77~

whole blood can be effectively removed by adsorp-tion.
As described above, the adsorber module and the adsorber apparatus of the present invention is very effective for the treatment of whole blood, but they also can be effectively used for removing the malig-nant components of plasma separated from whole blood by means of a conventional plasma separator.
Detailed Description of The Preferred Embodiments The present invention will now be described in more detail with reference to the following Examples, which should not be construed to be limit-ing the scope of the present invention.
Example 1 A high-density polyethylene (HI-ZEX 2208J, a product of Mitsui Petrochemical Co., Japan) having a density of 0.968 g/cm3 and a melt index, as measured in accordance with ASTM D1238, of 5.5 is extruded from an annular hollow fiber spinning nozzle having an annular orifice outside diameter of 35 mm and an annular orifice inside diameter of 27 mm (sllt width: 4 mm) at a winding temperature of 150 C and at a polymer extrusion rate of 16 g/min and a wind-ing rate of 200 m/min. The thus obtained hollow fiber is subjected to annealing at 115 C for 2 ~, , .
,, :: :: , .
~: :

~ ~325770 hours. The anneal~d hollow fiber is then cold-stretched at room temperature at a stretchlng ratio (times) (the ratio of the length of the stretched hollow fiber to the length of the hollow fiber before stretching, expressed by times) of 1.33 times by passing the annealed hollow fiber through stretching rolls arranged to provide a stretching path of 200 mm. Then, ~he cold-stretched hollow fiber is hot-stretched successively at 78 C, 95 C
and 98 C at stretching ratios at 78 C, 95 C and 98 C of 3 times, 1.28 times and 1.14 times, respec-tively, The ratio (~) of the length of the stretched hollow fiber to the original length of the , hollow fiber before cold-stretching and hot-stretch~
ing is 480 %, The thus stretched hollow fiber is heat set at 115 C for 2 min, to thereby obtain a porous polyethylene hollow fiber.
The polyethylene hollow iber is immersed in a ` solution of polyethylene vinyl alcohol (Soanol Z, manufactured and sold by The Nippon Synthetic Chemi-cals Industry Co., Ltd., Japan~ in 70 % (w/w) aque-ous ethanol solution, having a polyethylene vinyl alcohol concentration of 0~9 % by weightt and kept `~ at 50 C for 5 min in the solution. Then, the hol-`i ~5 low fiber is taken out of the solution and dried at ,.,~
; * Trademark :
_ 41 -'`

.'. ~' .: , , . . : : : - : :

~ 132577~

55 C for 1O5 hours. The resultant hollow fiber has an inside diameter of 340 llm, an outside diameter of 440 ~m, a membrane thlckness of 50 ~m, an average pore diameter of 0.3 ~m and a sur~ace area of 21 m2/g.
The hollow fiber is cut to the length of 30 cm to obtain 2000 cut hollow fibers, and they are im-mersed in a mixture of 500 ml of acetone, 390 ml of epichlorohydrin and 90 ml of a 40 ~ (w/w) aqueous NaOH solution. The hollow fibers are subjected to ultrasonication in the mixture at 30 C for 5 hours.
Thereafter, the hollow fibers are washed with ace-tone and then with water, to obtain epoxy-activated polyethylene hollow fibers.
The epoxy-activated polyethylene hollow fibers are immersed in 1000 ml of 1 M sodium carbonate buffer (pH 9.8) containing 10.20 g of tryptophan and sub~ected to ultrasonication at 50 C for 24 hours, to thereby link tryptophan to the overall surface of each of the hollow fibers. The amount of tryptophan linked as a ligand to the surface of each of the hollow fibers is 360 ~mol/g (dry basis). The ligand-linked hollow fibers are dried. The dried hollow fibers are inserted in a polyethylene-made cylindrical casing so that they are arranged sub-,, .
_ 42 -. : - -~ 1~2577~

stantially in parallel relationship and accommodated in the casing along the length thereof. Then, both end portions of the hollow fibers and both end por-tions of the inner side wall of a polycarbonate-made cylindrical casing having a plasma outlet on its side wall are bonded by a centrifugal mold~ng method using a polyurethane resin adhesive to obtain an assembly. Both the end portions of the resultant assembly are cut o~ to open the terminals of the hollow fibersl and an opening-having end cap is then attached to each of the end portions of the assembly as shown in Fig. 1 to provide a blood introduction means and a blood withdrawal means so that the both end portions of the hollow fibers are fluid-tightly connected to the blood introduction means and the blood withdrawal means, respectively, thereby estab-lishing communication between the blood introduction means and the blood withdrawal means through the hollow fibers. Thus, there is obtained an adsorber module as shown in Fig. 1. The average effective length o~ the hollow fibers of the adsorber module is 255 mm, and the ratio of the effective length of the hollow fiber to the square of the inside diame-ter of the hollow fiber (L/D2) = 2205 mm~1.
Using the adsorber module, an adsorber appara-. .

_ 43 -,~
.. .

.: , , , . ; . ~, .

~ ~325770 tus for whole blood treatment as shown in Fig. 4 is constructed.
A blood from a patient suffering from rheuma-toid arthritis is heparinized. An aliquot of the blood is subjected to measurement of amounts of rheumatoid factor, immune complex and platelet. The measurement of the amount of rheumatoid ~actor is conducted by a passlve hemagglutination test method using a RAHA test kit (manufactured and sold by Fuji Zoki Pharmaceutical Co., Ltd., Japan), and the mea-surement of the amount o~ immune complex is con-ducted by a Raji Cell method as described in "Immune Complex Disease", 1st ed., published by Ishiyaku Publishers, Inc., Tokyo, Japan (1982), pp. 93-94.
The measurement of the amount of platelet is con-ducted by a well known Brecher-Cronkite method in which a blood sample is diluted 100 times with a 1 %
aqueous ammonium oxalate solution and subjected to measurement of the platelet concentration by using a hemocytometer. The blood is lntroduced to the adsorber apparatus from the patient ~donor) through blood inlet 9 and flowed to adsorber module 13 at a flow rate of 50 ml/min through conduits 10 and 12 by means of pump 11 (blood transport means). In ad-sorber module 13, the blood is allowed to flow ..

. . . ~ .
.. . .... .

:........ . ,,.~ , ... . . ... .

2~77~

through the hollows of hollow fibers 1, and the plasma of the blood is allowed to pass through the membrane of each hollow fiber. The thus separated plasma is flowed to conduit 20 through a bypass, i.e., through conduits 16 and 18 by means of pump 17 at a flow rate of one third of the flow rate of the blood flowed into adsorber module 13 by means of pump 11. On the other hand, the cell-enriched blood which has not been passed through the bypass is flowed to conduit 20 through conduit 19. In conduit 20, the plasma is combined with the cell-enriched blood and the resultant mixture is returned to the donor through blood outlet 15. The blood from the donor is recycled through the same adsorber appara-tus. The recycling is continued for 30 min. With this adsorber apparatus, recycling of the blood is stably, sufficiently performed using an extracor-poreal circuit blood volume as small as 210 ml.
This volume is satisfactorily small from the view-point of attaining the whole blood treatment with a decreased extracorporeal circuit blood volume being taken outside the body.
The blood treatment by the adsorber apparatus can be stably conducted without blood coagulation and hemolysis.

~ ~ 32~77~

After the blood recycling for 30 min, an aliquot of the recycled blood is collected from blood outlet 20 and the amounts of rheumatoid factor, immune complex and platelet in the blood collected are measured in the same manner as mentioned above~
As a result, it is found that the amounts of the rheumatoid arthritis and immune complex befoxe the blood treatment which are 1280 and 120 ~g/ml, respectively, are decreased to 320 and 28 ~g/ml, respectively, after the blood treatment.
On the other hand, the platelet concentration before the treatment is 340,000 cells/mm3, whlle the platelet concentration a~ter the treatment is 300,000 cells/mm3. That is, the platelet concentra tion is not so lowered by the whole blood treatment.
As apparent from the above, by the use of the adsorber apparatus of the present invention, malig-nant substances ~i.e., rheumatoid factor and immune complex~ are selectively removed with little loss of plateletsO
Example 2 ~ Epoxy-activated hollow fibers are prepared in `s the same manner as in Example 1, and immersed in a , 25 30 % (w/w) aqueous dextran sulfate solution (pH 131 :~
~, _ 46 -' .

, :. - . . . .. : . , :: . :: . , : :

; ~ .

~ 132~77~

and kept at 50 C for 24 hours, to thereby link dextran sulfate (a ligand) to the overall sur~ace of each hollow fiber. The resultant hollow fibers are sufficiently washed with water. Uslng the washed S hollow fibers, an adsorber module is prepared in the same manner as in Example 1.
Then, using the thus obtained adsorber module, an adsorber apparatus for blood treatment is con-structed as shown in Fig. 3.
A blood from a patient suffering from familial hypercholesterolemia and having a hematocrit ~Ht) of 35 % is heparinized. An aliquot of the blood is subjected to measurement of the total cholesterol concentration and platelet concentration~ The measurement of total cholesterol concentration is conducted by an enzyme assay using a test kit Cholesterol C-Test Wako, manufactured and sold by Wako Pure Chemicals Industries Ltd., Japan. The most part of cholesterol present in the blood of a patient of familial hypercholesterolemia is derived ~rom low-density lipoproteins, and the term ~Itotal cholesterol" means to include cholesterol and low-density lipoproteins. The measurement of platelet concentration is conducted in the same manner as in Example 1. The blood is introduced to the adsorber .

~ - ~7 -: .: . : ~ :: :; . :

. ~ . .

~ ~32~770 apparatus from the patient (donor) through blood inlet 9 and flowed to adsorber module 13 at a flow rate of 50 ml/min through conduits 10 and 12 by means of pump 11. 10 Minutes later, it is observed that the interstices extending between the casing and the hollow fibers of the adsorber module and between the outer surfaces of individual hollow fibers have been filled with plasma assuming light yellow. The blood treated with the adsorber module i5 returned to the donor through blood outlet 15.
The blood from the donor is recycled through the same absorber apparatus in the same manner as in Example 1. The recycling is continued for 30 minutes. With this adsorber apparatus~ recycling of the blood is stably, suf~iciently performed using an extracorporeal circuit blood volume as small as 160 ml. This volume is satisfactorily small from the viewpoint of attaining the who~e blood treatment with a decreased extracorporeal circuit blood volume being taken outside the body. 30 Minutes later, an aliquot of the recycled blood is collected from blood outlet 15 and the total cholesterol concentra-tion of the collected blood is measured according to the same method as mentioned above. As a result, it ~, 25 is found that the total cholesterol concentration is , .
''~

.. . . .

~ ~32~7~

decreased to 110 mg/dl after the blood treatment, which is extremely low as compared to the total cholesterol concentration of the blood before the blood treatment, that is, 540 mg/dl~
On the other hand, the platelet concentration of the blood is also determined after the blood treatment. ~s a result, it is found that the plate-let concentration after the blood treatment is 250,000 cells/mm3, while the platelet concentration before the blood treatment is 270,000 cells/mm3.
That is, the platelet concentration is not so low~
ered by the blood treatment.
As apparent from the above, by the use of the adsorber apparatus of the present invention, a malignant substances ~i.e., cholesterol and low-density lipoproteins) are selectively removed with little loss of platelets.
Examples 3 to 5 A high-density polyethylene (HI-ZEX 2208J, a product of Mitsui Petrochemical Co., Japan~ having a d~nsity of 0.968 g/cm3 and a melt index, as measured in accordance with ASTM D1238, of 5~5 is extruded :( through a spinniny nozzle having an annular orifice . outside diameter of 33 mm and an annular orifice ;s 25 inside diameter of 25 mm at a winding temperature .

~ _ 49 -.

, ~ ~32~7~

of 150 C, and at a polymer extrusion rate of 16 g/min and a windlng rate as shown in Table 1.
The thus obtained hollow fiber is subjected to an-nealing at 115 C for 2 hours. The annealed hollow fiber is then cold-stretched at room temperature at a~stretching ratio (times) ~the ratio of the length of the stretched hollow fiber to the length of the hollow fiber before stretching, expressed by times) of 1.33 times by passing the hollow fiber through stretching rolls arranged to provide a stretching-path of 200 mm. Then, the cold-stretched hollow fiber is hot-stretched successively at 78 C, 95 C
and 98 C to thereby stretch the hollow fiber at a ratio (~) of the length of the hollow fiber to the original length of the hollow fiber before cold-stretching and hot-stretching, of 480 % The resul-tant hollow fiber is heat set at 115 C for 2 min, to thereby obtain a porous polyethylene hollow fiber.
The polyethylene hollow fiber is subjected successively to polyvinyl alchohol-treatment, epoxy-activation, and treatment for linking ligands in the same manner as in Example 1. Then, an adsorber module for blood treatment is prepared using the porous polyethylene hollow fiber in the same manner ;'~ .

k~' ~3257 7 ~

as in Example 1. The average inside diameter (D), the average effective length (L) and the total surface area of the hollow fibers in the adsorber module are shown in Table 1. Using the adsorber module, substantially the same adsorber apparatus for blood treatment as shown in Fig. 4 except that a plasma circuit including pump 17 and conduit 18 is removed, and a container is attached to conduit 16 in order to collect plasma separated by adsorber module 13 is constru~ted.
A blood from a patient (donor~ suffering from rheumatoid arthritis is heparinized and introduced to the adsorber apparatus through blood inlet 9.
The introduced blood is flowed to adsorber module 13 through conduits 10 and 12 by means of pump 11, and plasma separated by adsorber module 13 is collected through plasma outlet 16. With this adsorber appa-ratus, recycling of the blood is stably, sufficient-ly performed using an extracorporeal clrcuit blood volume as small as 210 ml. This volume is satis-factorily small from the viewpoint of attaining the whole blood treatment with a decreased extracor-poreal circuit blood volume being taken outside the body. The amount of the collected plasma is mea-., sured. Further, the rheumatoid factor concentration ' : :

. , .

~3%~7o of the collected plasma is determined in the same manner as in Example 1. The results are shown in Table 1.
As apparent from the results shown in ~able 1, although the average inside diameters of the indivi-dual hollow fibers are different between the hollow fibers used in Examples 3 to 5y the surface areas of the hollow fibers in the prepared adsorber modules of Examples 3 to 5 are almost the same~ However, the larger the L/D2 value, the smaller the amount of rheumatoid factor in the treated blood becomes. On the other hand, the larger the L/D2 value, the larger the collection rate of plasma becomes. This means that the smaller the inside diameter of the hollow fiber, the higher the efficiency of plasma separation becomes. Particularly, when the L/D2 value is 2,000 mm~1 or higher as in Examples 3 and 4, plasma separation can be conducted with extremely high efficiency.

:
~;

:: . : . ~

~ ~257~

o $ ~ oO ~ o o o h ~1 ~ ~ o~
~ Q~ ~1 ~ 4~
o o o o a) o h '~J ~ ~
~ ~ .
1~ ~ ~ ~ ~: o h~ ~ ~ ,1 . co O ~ . ~1 u~ h ~:4-- O u~
~:

O O 3 11~ I h a) _ o . o o ~
~ O h r~l a) ~ ~ o ~ (d 4 1 ~ ~1 a) ,a Q ~ ~3 ~1 ,1 ~J ~I) h O ~J R h ~ -- . _ h ~A h~ ~ 0 ~
,~ ~0 ,1 ~ o o o -- R
R ~ 1~ o o o .~j ,1:;

o 4~
~41 U~ A

1~ 0 ~ O h ~ o o o ~rl h h O t~ ~1 0 t~l ~ ~`I a~ ~
a) ~ ~ t) 0-~ ~ 0 -- h 4~
O rl h 30 r~l O
r-l ~ O O O ~ E~
~a a) ~ h a ~ ~ ~ O
A ` o o o ~1 ~
m h ~_ ..
~^ ~o r¦ rl O O O rl ~ E~; o o o r ~ h-- O
~a~ , ~ ~r ~

~ P~ .

. ; ': :

~ ~325770 Comparative Example 1 An adsorber material comprising a granular carrier and tryptophan linked to the carrier is used for the adsorption of rheumatoid factor. The ad-sorber material is produced as follows.
A uniform mixture of 100 g of vinyl acetate, 64.3 g of triallyl isocyanurate, 100 g of ethyl acetate, 100 g of heptane, 7.5 g of polyvinyl ace-tate (deyree of polymerization: 500) and 3.8 g of 2,2'-azobisisobutyronitrile, and 4G0 ml of an aque-ous solution containing 1 % by weight of polyvinyl alcohol, 0.05 % by weight of sodium dihydrogenphos-phate dihydrate and 1.5 % by weight of sodium di-hydrogenphosphate dodecahydrate are charged in a flask and sufficiently mixed by stirring. Then, the resultant mixture is heated, while stirring, suc-cessively at 65 C for 18 hours and at 75 C for 5 hours to conduct a suspension polymerization. The thus obtained granular copolymer is filtered off and washed with water. The thus obtained copolymer is subjected to acetone extraction. The resultant copolymer is subjected to ester exchange reaction at 40 C for 18 hours in a solution of 46.S g of sodlum hydroxide in 2 Q of methanol. The thus obtained gel of the copolymer has an average particle diameter of .

.
- :

100 ~m, a vlnyl alcohol unit (qOH) per unit weight-of 8.9 meq/g, a specific surface area of 65 m2/g and a cut-off molecular weight of 9 x105 as measured using a dextran.
Then, 50 g of the obtained gel (dry basis) is suspended in 600 ml of dimethyl sul~oxide and to the resultant suspension are added 391.5 ml o~ epichlo-rohydrin and 50 ml of 30 % (w/w) aqueous sodium hydroxide solution. The mixture is stirred at 30 C
for 5 hours to activate the gel~
After the gel activation, the resultant gel is washed successively with dimethylsulfoxide and with water, and suction-dehydrated. The thus obtained activated yel is suspended in 800 ml of 0.1 M sodium carbonate buffer containing 8.15 g of tryptophanO
The suspension is stlrred at 50 C for 14 hours to link the tryptophan to the overall surface of the gel. Thereafter, to the mixture is added 165 ml oE
an aqueous tris~hydroxyethyl)aminomethane solution having a tris(hydroxyethyl)aminomethane concentra-tion of 60.6 mg/ml. Then, the resultant mixture is heated at 50 C for 5 hours while stirring so that the active groups of the gel remainlng unlinked by tryptophan are blocked. The resultant gel is suffi-ciently washed with water, to thereby obtain an .: . . . .
.., .. ~ , .

"` . ` ' ~, , ,. :

X ~ 32~770 adsorber material. The amount of the tryptophan linked to the adsorber material is 360 ~mol/g (dry basis).
The adsorber material is filled in a column having an inside diameter of 40 mm and a length of 318 mm to obtain an adsorber column. Using the adsorber column, an adsorber apparatus as shown in Fig. 5 is constructed. In Fig. 5, numeral 24 desig-nates the adsorber column and numeral 21 designates 1~ a plasma separator. The adsorber column 24 is con-nected to the plasma outlet o plasma separator 21 through conduit 21, pump 17 and conduit 23 so that the plasma separated by plasma separator 21 can be conveyed to adsorber column 24 by means of pump 17.
As the plasma separator, Plasmaflo~ AP05H manufac-tured and sold by Asahi Medical Co., Ltd., Japan is used.
A blood from a patient suffering from rheuma-toid arthrltis is heparinized. An aliquot of the blood is subjected to measurement of amount of rheumatoid factor by a passive hemagglutination test method using a RAHA test kit Imanufactured and sold by Fuji Zoki Pharmaceutical Co., Ltd., Japan). The blood is introduced to the adsorber apparatus from the patient (donor) through blood inlet 9 and flowed ~32577~

to plasma separator 21 at a flow rate o~ 50 ml/min through conduits 10 and 12 by means of pump 11. The blood is separated into plasma and cell-enriched blood by plasma separator 21, and the plasma is flowed to adsorber column 24 through conduits 22 and 23 by means of pump 17 at a flow rate oE 17 m/min.
In conduit 27, the plasma treated with adsorber column 24 and flowed through conduit 25 is combined with cell-enriched blood flowed through conduit 26, and the resultant mixture is returned to the donor through blood outlet 15~ The blood from the donor is recycled through the same adsorber apparatus.
The recycling is continued for 30 min.
After the blood recycling for 30 min, an lS aliquot of the recycled blood is collected from blood outlet 27 and the amount o~ rheumatoid factor in the blood is measured in the same manner as in Example 1.
As a result, it is ~ound that the amount of rheumatoid factor before the blood treatment which is 1280, is decreased to 160 after the blood treat-ment, which is comparable to the amount of the rheumatoid factor in the blood after the blood treatment conducted in Example 3, Howe~er, with this adsorber apparatus, recycling o~ the blood is :' ~ - 57 -... .

, - ., - . .
.:: . . . , i . . ~ :

132~77~

performed using an extracorporeal circuit blood volume as large as 610 ml. This volume is disadvan-tageously very large so that a heavy burden is posed on the patient during the blood treatment.

( ..

., - , .

Claims (14)

1.
What is claimed is:
1. An adsorber module for whole blood treatment com-prising:
a casing provided with a blood introduction means and a blood withdrawal means, and a plurality of porous hollow fibers substantially equal in length arranged substantially in parallel rela-tionship and bonded together at both end portions thereof to form a bundle, each porous hollow fiber of said bundle having openings at both terminal ends thereof, said bundle being disposed in said casing along the length of said casing, said both end portions of the hollow fibers of said bundle being fluid-tightly connected to said blood introduc-tion means and said blood withdrawal means, respectively, thereby establishing communication between said blood introduction means and said blood withdrawal means through said bundle of hollow fibers, each porous hollow fiber comprising a membranous porous resin matrix having pores which open at least at the inner wall of the hollow fiber and a plurality of ligands attached to the overall surface of said porous resin matrix, said ligands being capable of interacting with a

2.
substance to be adsorbed.

2. The module according to claim 1, wherein said ligand has low antigenicity.

3. The module according to claim 1, wherein said ligand is linked to the overall surface of said porous resin matrix by covalent bonding.

4. The module according to claim 1, wherein said ligand is a hydrophobic compound.

5. The module according to claim 1, wherein said ligand is a polyanion.

6. The module according to claim 1, wherein said porous resin matrix has an average pore diameter of from 0.005 to 3 µm.

7. The module according to claim 6, wherein said average pore diameter is in the range of from 0.01 to 2 µm.

8. The module according to claim 1, wherein said membranous porous resin matrix of each hollow fiber has pores therewithin and openings on both surfaces thereof, 3.
said pores cooperating with said openings to form through-paths running between both the surfaces of said resin matrix.

9. The module according to claim 1, wherein said porous hollow fibers have an average effective length (L mm) and an average inner diameter (D mm) which satisfy the inequal-ity:
L/D2 (mm-1) ? 2000 said average effective length being defined as an average of the lengths of said porous hollow fibers minus the lengths of said both end portions of said porous hollow fibers at which the fibers are bonded together and fluid-tightly connected to said blood introduction means and said blood withdrawal means, respectively.

10. The module according to claim 8, wherein said casing is further provided with at least one opening for withdraw-ing the plasma.

11. The module according to claim 1, wherein said resin matrix is composed of at least one resin selected from the group consisting of cellulose, a cellulose derivative, a water-insoluble polyvinyl alcohol, a copolymer of ethylene and vinyl alcohol, a polyolefin, a polysulfone and a poly-4.

tetrafluoroethylene.

12. An adsorber apparatus for whole blood treatment com-prising:
(a) an adsorber module comprising:
a casing provided with a blood introduction means and a blood withdrawal means, and a plurality of porous hollow fibers substantially equal in length arranged substantially in parallel rela-tionship and bonded together at both end portions thereof to form a bundle, each porous hollow fiber of said bundle having openings at both terminal ends thereof, said bundle being disposed in said casing along the length of said casing, said both end portions of the hollow fibers of said bundle being fluid-tightly connected to said blood introduc-tion means and said blood withdrawal means, respectively, thereby establishing communication between said blood introduction means and said blood withdrawal means through said bundle of hollow fibers, each porous hollow fiber comprising a membranous porous resin matrix having pores which open at least at the inner wall of the hollow fiber and a plurality of ligands attached to the overall surface of said porous resin matrix, 5.

said ligands being capable of interacting with a substance to be adsorbed, (b) a blood introducing passage means comprising a first conduit having one end fluid-tightly connected to said blood introduction means of module (a), a second con-duit having a blood inlet at one end thereof, and a blood transport means disposed between and fluid-tightly con-nected to the other ends of said first and second conduits and (c) a blood withdrawing passage means comprising a third conduit having one end fluid-tightly connected to said blood withdrawal means of module (a) and having a blood outlet at the other end thereof.

13. The apparatus according to claim 12, wherein said membranous porous resin matrix of each hollow fiber of said adsorber module has pores therewithin and openings on both surfaces thereof, said pores cooperating with said opening to form through-paths running between both the surfaces of said resin matrix.

14. The apparatus according to claim 13, wherein said casing of module (a) is further provided with at least one opening for withdrawing the plasma, said opening being fluid-tightly connected to said 6.
blood withdrawing passage means through a bypass comprising a fourth conduit having one end fluid-tightly connected to said opening, a fifth conduit having one end fluid-tightly connected to said blood withdrawing passage at its inter-mediate position, and a plasma transport means disposed between and fluid-tightly connected to the other ends of said fourth and fifth conduits.